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Literature summary for 5.1.2.2 extracted from

  • Bourque, J.R.; Bearne, S.L.
    Mutational analysis of the active site flap (20s loop) of mandelate racemase (2008), Biochemistry, 47, 566-578.
    View publication on PubMed
Search for more literature for 5.1.2.2

Cloned(Commentary)

Cloned (Comment) Organism
Recombinant MR from is overproduced from Escherichia coli strain BL21(DE3) cells transformed with a pET-15b plasmid containing the MR open reading frame. This construct encodes the MR gene product with an N-terminal hexahistidine tag. Mutants are produced by site-directed mutagenesis Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
A25V Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
A25V variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
T24S Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
T24S variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V22A Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V22A variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V22F Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V22F variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V22I Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V22I variation of the hydrophobic loop. For mandelate, catalytic efficiency similar to wild-type. For substrate 2-naphthylglycolate, increase in catalytic efficiency Pseudomonas putida
V22I/V29L Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V22I/V29L variation of the hydrophobic loop, catalytic efficiency similar to wild-type Pseudomonas putida
V26A Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V26A variation of the hydrophobic loop, catalytic efficiency similar to wild-type Pseudomonas putida
V26A/V29L Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V26A/V29L variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V26F Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V26F variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V26L Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V26L variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V29A Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V29A variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V29F Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V29F variation of the hydrophobic loop, decrease in catalytic efficiency Pseudomonas putida
V29L Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate Pseudomonas putida
V29L variation of the hydrophobic loop. For mandelate, catalytic efficiency similar to wild-type. For substrate 2-naphthylglycolate, increase in catalytic efficiency Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
2-naphthohydroxamate competitive inhibition at pH 7.5 Pseudomonas putida
2-naphtholhydroxamate
-
 Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
 Mg2+ pH 7.5, mutant V22A, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.04
-
 Mg2+ pH 7.5, mutant V22F, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.05
-
 Mg2+ pH 7.5, mutant V26F, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.08
-
 Mg2+ pH 7.5, mutant V29A, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.12
-
 Mg2+ pH 7.5, mutant V26L, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.15
-
 Mg2+ pH 7.5, mutant V26A, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.22
-
 Mg2+ pH 7.5, mutant V29F, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.29
-
 Mg2+ pH 7.5, mutant T24S, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.34
-
 Mg2+ pH 7.5, wild-type, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.47
-
 Mg2+ pH 7.5, mutant V29L, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.51
-
 Mg2+ pH 7.5, mutant V26A/V29L, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.55
-
 (S)-2-naphthylglycolate mutant V29F, pH 7.5, 25°C Pseudomonas putida
0.55
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
0.6
-
 (S)-Mandelate mutant V29L, pH 7.5, 25°C Pseudomonas putida
0.6
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
0.62
-
 (S)-2-naphthylglycolate mutant V22F, pH 7.5, 25°C Pseudomonas putida
0.62
-
 (S)-2-naphthylglycolate mutant V26L, pH 7.5, 25°C Pseudomonas putida
0.62
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
0.62
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
0.64
-
 Mg2+ pH 7.5, mutant A25V, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
0.74
-
 (S)-2-naphthylglycolate mutant V26A, pH 7.5, 25°C Pseudomonas putida
0.74
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
0.8
-
 (S)-2-naphthylglycolate mutant V29L, pH 7.5, 25°C Pseudomonas putida
0.8
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
0.9
-
 (R)-mandelate mutant V29F, pH 7.5, 25°C Pseudomonas putida
0.9
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
0.91
-
 (R)-mandelate mutant V26A, pH 7.5, 25°C Pseudomonas putida
0.91
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
0.93
-
 (S)-2-naphthylglycolate mutant V29A, pH 7.5, 25°C Pseudomonas putida
0.93
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
0.97
-
 (S)-Mandelate wild-type, pH 7.5, 25°C Pseudomonas putida
0.97
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type Pseudomonas putida
1
-
 (S)-2-naphthylglycolate wild-type, pH 7.5, 25°C Pseudomonas putida
1
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type Pseudomonas putida
1.07
-
 (S)-Mandelate pH 7.5 at 25°C, 20 mM Mg2+, wild-type Pseudomonas putida
1.07
-
 (S)-Mandelate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1.1
-
 (R)-mandelate mutant A25V, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (S)-Mandelate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (S)-Mandelate mutant V26F, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (S)-2-naphthylglycolate mutant V26F, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (S)-Mandelate mutant V26L, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (S)-Mandelate mutant V29F, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (R)-2-naphthylglycolate mutant V29F, pH 7.5, 25°C Pseudomonas putida
1.1
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
1.1
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
1.1
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
1.1
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
1.1
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
1.1
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
1.1
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
1.2
-
 (R)-mandelate wild-type, pH 7.5, 25°C Pseudomonas putida
1.2
-
 (R)-2-naphthylglycolate wild-type, pH 7.5, 25°C Pseudomonas putida
1.2
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type Pseudomonas putida
1.2
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type Pseudomonas putida
1.3
-
 (R)-2-naphthylglycolate mutant V26L, pH 7.5, 25°C Pseudomonas putida
1.3
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
1.4
-
 (S)-2-naphthylglycolate mutant T24S, pH 7.5, 25°C Pseudomonas putida
1.4
-
 (R)-mandelate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
1.4
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
1.4
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
1.47
-
 (R)-mandelate pH 7.5 at 25°C, 20 mM Mg2+, wild-type Pseudomonas putida
1.47
-
 (R)-mandelate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1.5
-
 (S)-Mandelate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1.5
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
1.55
-
 (S)-2-naphthylglycolate mutant V22A, pH 7.5, 25°C Pseudomonas putida
1.55
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
1.6
-
 (R)-2-naphthylglycolate mutant V29L, pH 7.5, 25°C Pseudomonas putida
1.6
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 20 mM Mg2+, wild-type Pseudomonas putida
1.6
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
1.6
-
 (S)-2-naphthylglycolate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1.67
-
 (S)-2-naphthylglycolate mutant A25V, pH 7.5, 25°C Pseudomonas putida
1.67
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
1.7
-
 (S)-Mandelate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1.7
-
 (S)-Mandelate mutant V26A, pH 7.5, 25°C Pseudomonas putida
1.7
-
 (S)-2-naphthylglycolate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
1.7
-
 (R)-2-naphthylglycolate mutant V26F, pH 7.5, 25°C Pseudomonas putida
1.7
-
 (R)-mandelate mutant V26L, pH 7.5, 25°C Pseudomonas putida
1.7
-
 (R)-mandelate mutant V29L, pH 7.5, 25°C Pseudomonas putida
1.7
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
1.7
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
1.7
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
1.7
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
1.7
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
1.7
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
1.8
-
 (S)-Mandelate mutant A25V, pH 7.5, 25°C Pseudomonas putida
1.8
-
 (R)-mandelate mutant V26F, pH 7.5, 25°C Pseudomonas putida
1.8
-
 (R)-2-naphthylglycolate mutant V29A, pH 7.5, 25°C Pseudomonas putida
1.8
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
1.8
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
1.8
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
1.9
-
 (R)-2-naphthylglycolate mutant A25V, pH 7.5, 25°C Pseudomonas putida
1.9
-
 (R)-2-naphthylglycolate mutant V22A, pH 7.5, 25°C Pseudomonas putida
1.9
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
1.9
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
2.1
-
 (R)-2-naphthylglycolate mutant V26A, pH 7.5, 25°C Pseudomonas putida
2.1
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
2.2
-
 (R)-2-naphthylglycolate mutant V22F, pH 7.5, 25°C Pseudomonas putida
2.2
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
2.4
-
 (S)-2-naphthylglycolate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
2.4
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
2.8
-
 (R)-mandelate mutant T24S, pH 7.5, 25°C Pseudomonas putida
2.8
-
 (R)-mandelate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
2.8
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
2.8
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I/V29L Pseudomonas putida
2.9
-
 (R)-mandelate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
2.9
-
 (S)-2-naphthylglycolate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
2.9
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
2.9
-
 (S)-2-naphthylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
3
-
 (S)-Mandelate mutant T24S, pH 7.5, 25°C Pseudomonas putida
3
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
3.2
-
 (R)-2-naphthylglycolate mutant T24S, pH 7.5, 25°C Pseudomonas putida
3.2
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
3.4
-
 (S)-Mandelate mutant V22A, pH 7.5, 25°C Pseudomonas putida
3.4
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
3.5
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 20 mM Mg2+, wild-type Pseudomonas putida
3.5
-
 (R)-2-naphthylglycolate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
3.7
-
 (R)-2-naphthylglycolate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
3.7
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I/V29L Pseudomonas putida
3.8
-
 Mg2+ pH 7.5, mutant V22I, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
3.9
-
 (S)-Mandelate mutant V29A, pH 7.5, 25°C Pseudomonas putida
3.9
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
4.2
-
 (R)-2-naphthylglycolate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
4.2
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
4.2
-
 Mg2+ pH 7.5, mutant V22I/V29L, addition of either 10 mM (R)- or (S)-mandelate Pseudomonas putida
4.4
-
 (R)-mandelate mutant V22A, pH 7.5, 25°C Pseudomonas putida
4.4
-
 (R)-mandelate mutant V22F, pH 7.5, 25°C Pseudomonas putida
4.4
-
 (R)-2-naphthylglycolate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
4.4
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
4.4
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
4.4
-
 (R)-2-naphtylglycolate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
5
-
 (R)-mandelate mutant V29A, pH 7.5, 25°C Pseudomonas putida
5
-
 (R)-mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
6.1
-
 (S)-Mandelate mutant V22F, pH 7.5, 25°C Pseudomonas putida
6.1
-
 (S)-Mandelate pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ catalysis is Mg2+-dependent Pseudomonas putida
Mg2+ Km value for wild-type, 0.34 mM Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40700
-
 mutants V22A, V26A and V29A Pseudomonas putida
40710
-
 mutants V26A/V26L and T24S Pseudomonas putida
40730
-
 wild type Pseudomonas putida
40740
-
 mutants V22I, V26L and V29L Pseudomonas putida
40760
-
 mutants A25V and V22I/V29L Pseudomonas putida
40780
-
 mutants V22F, V26F and V29F Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-mandelate Pseudomonas putida catalyzes the Mg2+-dependent 1,1-proton transfer that interconverts the enantiomers, catalysis is done via a two-base mechanism (R)-mandelate
-
 r

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P11444
-
-

Purification (Commentary)

Purification (Comment) Organism
by metal ion affinity chromatography Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-2-naphthylglycolate
-
 Pseudomonas putida (S)-2-naphthylglycolate
-
 r
(R)-2-naphtylglycolate
-
 Pseudomonas putida (S)-2-naphthylglycolate
-
 r
(R)-mandelate
-
 Pseudomonas putida (S)-mandelate
-
 r
(S)-2-naphthylglycolate
-
 Pseudomonas putida (R)-2-naphthylglycolate
-
 r
(S)-2-naphthylglycolate racemisation Pseudomonas putida (R)-2-naphthylglycolate
-
 r
(S)-mandelate
-
 Pseudomonas putida (R)-mandelate
-
 r
(S)-mandelate catalyzes the Mg2+-dependent 1,1-proton transfer that interconverts the enantiomers, catalysis is done via a two-base mechanism Pseudomonas putida (R)-mandelate
-
 r

Synonyms

Synonyms Comment Organism
mandelate racemase
-
 Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.95
-
 (S)-2-naphthylglycolate mutant V22F, pH 7.5, 25°C Pseudomonas putida
2.95
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
3 6 (R)-mandelate mutant V26L, pH 7.5, 25°C Pseudomonas putida
3 6 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
5.4
-
 (R)-2-naphthylglycolate mutant A25V, pH 7.5, 25°C Pseudomonas putida
5.4
-
 (S)-2-naphthylglycolate mutant V26F, pH 7.5, 25°C Pseudomonas putida
5.4
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
5.4
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
5.8
-
 (S)-2-naphthylglycolate mutant A25V, pH 7.5, 25°C Pseudomonas putida
5.8
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
7.1
-
 (S)-2-naphthylglycolate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
7.1
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
7.3
-
 (R)-2-naphthylglycolate mutant V26F, pH 7.5, 25°C Pseudomonas putida
7.3
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
8.2
-
 (S)-2-naphthylglycolate mutant V26L, pH 7.5, 25°C Pseudomonas putida
8.2
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
8.9
-
 (S)-2-naphthylglycolate mutant T24S, pH 7.5, 25°C Pseudomonas putida
8.9
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
9.7
-
 (S)-2-naphthylglycolate mutant V29F, pH 7.5, 25°C Pseudomonas putida
9.7
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
9.8
-
 (S)-2-naphthylglycolate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
9.8
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
10.4
-
 (R)-2-naphthylglycolate mutant V22F, pH 7.5, 25°C Pseudomonas putida
10.4
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
10.5
-
 (R)-2-naphthylglycolate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
10.5
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22I/V29L Pseudomonas putida
11.1
-
 (S)-2-naphthylglycolate mutant V22A, pH 7.5, 25°C Pseudomonas putida
11.1
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
13
-
 (R)-mandelate mutant A25V, pH 7.5, 25°C Pseudomonas putida
13
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
14
-
 (S)-2-naphthylglycolate mutant V26A, pH 7.5, 25°C Pseudomonas putida
14
-
 (R)-2-naphthylglycolate mutant V26L, pH 7.5, 25°C Pseudomonas putida
14
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
14
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
14.1
-
 (R)-2-naphthylglycolate mutant V29F, pH 7.5, 25°C Pseudomonas putida
14.1
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
15.6
-
 (R)-2-naphthylglycolate mutant V22A, pH 7.5, 25°C Pseudomonas putida
15.6
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
18.3
-
 (R)-2-naphthylglycolate mutant T24S, pH 7.5, 25°C Pseudomonas putida
18.3
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
22
-
 (S)-Mandelate mutant A25V, pH 7.5, 25°C Pseudomonas putida
22
-
 (S)-Mandelate mutant V26F, pH 7.5, 25°C Pseudomonas putida
22
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant A25V Pseudomonas putida
22
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
23
-
 (R)-2-naphthylglycolate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
23
-
 (S)-Mandelate mutant V26L, pH 7.5, 25°C Pseudomonas putida
23
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
23
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V26L Pseudomonas putida
30
-
 (R)-2-naphthylglycolate mutant V26A, pH 7.5, 25°C Pseudomonas putida
30
-
 (S)-2-naphthylglycolate mutant V29A, pH 7.5, 25°C Pseudomonas putida
30
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
30
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
32
-
 (S)-2-naphthylglycolate wild-type, pH 7.5, 25°C Pseudomonas putida
32
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, wild-type Pseudomonas putida
33
-
 (R)-mandelate mutant V26F, pH 7.5, 25°C Pseudomonas putida
33
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V26F Pseudomonas putida
38.4
-
 (S)-2-naphthylglycolate pH 7.5, 20 mM Mg2+, wild-type Pseudomonas putida
38.4
-
 (S)-2-naphthylglycolate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
46
-
 (R)-2-naphthylglycolate wild-type, pH 7.5, 25°C Pseudomonas putida
46
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, wild-type Pseudomonas putida
53
-
 (R)-2-naphthylglycolate mutant V29A, pH 7.5, 25°C Pseudomonas putida
53
-
 (S)-Mandelate mutant V29F, pH 7.5, 25°C Pseudomonas putida
53
-
 (R)-mandelate mutant V29F, pH 7.5, 25°C Pseudomonas putida
53
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
53
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
53
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V29F Pseudomonas putida
65
-
 (S)-2-naphthylglycolate mutant V29L, pH 7.5, 25°C Pseudomonas putida
65
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
70
-
 (R)-2-naphtylglycolate pH 7.5, 20 mM Mg2+, wild-type Pseudomonas putida
70
-
 (R)-2-naphthylglycolate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
77
-
 (S)-Mandelate mutant V22A, pH 7.5, 25°C Pseudomonas putida
77
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+ mutant V22A Pseudomonas putida
82
-
 (R)-mandelate mutant V22A, pH 7.5, 25°C Pseudomonas putida
82
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V22A Pseudomonas putida
89
-
 (S)-Mandelate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
89
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
98
-
 (S)-Mandelate mutant V22F, pH 7.5, 25°C Pseudomonas putida
98
-
 (R)-mandelate mutant V22F, pH 7.5, 25°C Pseudomonas putida
98
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
98
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V22F Pseudomonas putida
101
-
 (S)-2-naphthylglycolate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
101
-
 (S)-2-naphthylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
102
-
 (R)-mandelate mutant T24S, pH 7.5, 25°C Pseudomonas putida
102
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
106
-
 (R)-mandelate mutant V26A/V29L, pH 7.5, 25°C Pseudomonas putida
106
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
120
-
 (S)-Mandelate mutant T24S, pH 7.5, 25°C Pseudomonas putida
120
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant T24S Pseudomonas putida
137
-
 (R)-2-naphthylglycolate mutant V29L, pH 7.5, 25°C Pseudomonas putida
137
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
147
-
 (R)-2-naphthylglycolate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
147
-
 (R)-2-naphtylglycolate pH 7.5, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
176
-
 (S)-Mandelate mutant V29A, pH 7.5, 25°C Pseudomonas putida
176
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
180
-
 (S)-Mandelate mutant V29L, pH 7.5, 25°C Pseudomonas putida
180
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
240
-
 (R)-mandelate mutant V29A, pH 7.5, 25°C Pseudomonas putida
240
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V29A Pseudomonas putida
300
-
 (R)-mandelate mutant V29L, pH 7.5, 25°C Pseudomonas putida
300
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V29L Pseudomonas putida
304
-
 (R)-mandelate mutant V26A, pH 7.5, 25°C Pseudomonas putida
304
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
470
-
 (S)-Mandelate wild-type, pH 7.5, 25°C Pseudomonas putida
470
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, wild-type Pseudomonas putida
492
-
 (S)-Mandelate pH 7.5, 20 mM Mg2+, wild-type Pseudomonas putida
492
-
 (S)-Mandelate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
550
-
 (S)-Mandelate mutant V26A, pH 7.5, 25°C Pseudomonas putida
550
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V26A Pseudomonas putida
552
-
 (R)-mandelate wild-type, pH 7.5, 25°C Pseudomonas putida
552
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, wild-type Pseudomonas putida
620
-
 (R)-mandelate pH 7.5, 20 mM Mg2+, wild-type Pseudomonas putida
620
-
 (R)-mandelate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
710
-
 (S)-Mandelate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
710
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L Pseudomonas putida
740
-
 (S)-Mandelate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
740
-
 (S)-Mandelate pH 7.5, 3.3 mM Mg2+, mutant V22I Pseudomonas putida
1010
-
 (R)-mandelate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1010
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V22I/V29L Pseudomonas putida
1150
-
 (R)-mandelate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
1150
-
 (R)-mandelate pH 7.5, 3.3 mM Mg2+, mutant V22I Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Pseudomonas putida

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.023
-
 2-naphthohydroxamate wild-type, 20 mM Mg2+, pH 7.5 Pseudomonas putida
0.023
-
 2-naphtholhydroxamate wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
0.027
-
 2-naphtholhydroxamate wild-type, pH 7.5, 25°C Pseudomonas putida
0.027
-
 2-naphthohydroxamate wild-type, 3.3 mM Mg2+, pH 7.5 Pseudomonas putida
0.035
-
 2-naphthohydroxamate mutant V29L, 3.3 mM Mg2+, pH 7.5 Pseudomonas putida
0.035
-
 2-naphtholhydroxamate mutant V29L, pH 7.5, 25°C Pseudomonas putida
0.112
-
 2-naphtholhydroxamate mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida
0.112
-
 2-naphthohydroxamate mutant V29I/V29I, 20 mM Mg2+, pH 7.5 Pseudomonas putida
0.19
-
 2-naphthohydroxamate mutant V22I, 3.3 mM Mg2+, pH 7.5 Pseudomonas putida
0.19
-
 2-naphtholhydroxamate mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ Pseudomonas putida